SMYD2 CRISPR Activation Plasmid (h): sc-403129-ACT

SMYD2 encodes a SET and MYND domain–containing lysine methyltransferase that modulates chromatin state and protein function through mono- and di-methylation of histone and non-histone substrates. By influencing transcriptional programs, DNA damage responses, and cell-cycle control, SMYD2 integrates epigenetic regulation with signaling pathways that shape proliferation and cellular stress adaptation. Reported substrates include regulators of genome stability and checkpoint signaling, linking SMYD2 activity to oncogenic networks and altered differentiation states observed across multiple tumor contexts. Consequently, SMYD2 is widely studied as a node connecting chromatin remodeling, transcriptional control, and proteome methylation in disease-relevant biology.

SMYD2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SMYD2 expression without altering the underlying DNA sequence.

SMYD2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SMYD2 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the SMYD2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SMYD2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SMYD2 locus and enabling the study of SMYD2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SMYD2 pathway restoration in tumor cells with silenced or reduced SMYD2 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.